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1.
细胞分裂素的混合抗体型免疫亲和柱的制备与应用   总被引:2,自引:0,他引:2  
陈以峰  郑志富 《生物技术》1994,4(5):22-23,26
内源细胞分裂素可以分成三组:异戊烯基腺苷组(iPAs)、玉米素核苷组(ZRs)、二氢玉米素核苷组(DHZRs)。从这三组分别选iPA、ZR、DHZR为半抗原合成免疫原,获得的三种免抗血清基本上只识别相应组的细胞分裂素。将经初纯化的三种抗血清偶联到CNBr活化的Sepharose4B上制成免疫亲和柱。在其中的一根柱床高2.1cm、体积3.55ml、直径1.5cm的亲和柱上,在流速1.5ml/min、80%冷甲醇为洗脱剂的条件下测得该柱容量为3.6—4.0μg、回收率达93.9%~98.3%。用该柱对丝瓜茎木质部伤流中细胞分裂素进行了纯化.表明能够用此柱对植物粗提液中的细胞分裂素进行快速分离纯化。  相似文献   

2.
丹皮酚对心肌细胞自律性和延迟后除极的影响   总被引:8,自引:0,他引:8  
目的与方法:采用常规玻璃微电极技术研究丹皮酚对离体心肌细胞自律性(AM)、延迟后除极(DAD) 及触发活动(TA)的影响。结果:1.8×10-4mol/L丹皮酚灌流组,肾上腺素(Adr)的阈浓度空白对照组为(1.28±0.57)μmol/L,药后为(1.56±0.53)μmol/L(n=9,P>0.05);用(1.8×10- 3) mol/L丹皮酚(Pae)灌流组,Adr 浓度由空白对照组的(1.22 ±0.62)μmol/L升高到(6.22±2.11)μmol/L(n=9,P<0.01)。1.8×10-3mol/L的Pae 能明显抑制哇巴因(Oua)诱发的DAD的幅值,当基本刺激周长为500,400,300 和200 ms 时,其DAD幅值从(5.5±2.0)mV,(7.3±2.1)mV,(8.0 ±2.4)mV和(9.2±1.9)mV减小到(3.0±1.1)mV、(3.6±1.7)mV,(4.3±2.0) mV和(5.9 ±1.6) mV,P<0.01。当基本刺激周长为200 ms时,TA 数目由5.5±1.0 降至0.7±0.3(P<0.01)。结论:丹皮酚能抑制心肌细胞AM、DAD及TA,具有抗心律失常作用  相似文献   

3.
三种类型辐射对质粒超螺旋DNA损伤的研究   总被引:7,自引:0,他引:7  
用agarose电泳和图象处理技术比较了60Coγ射线、UV及低能N+离子处理pUC19DNA超螺旋结构的损伤效应及若干自由基清除剂的保护效应。结果表明:(1)γ射线和UV照射干燥DNA的损伤显著低于水溶液样品;(2)N+离子注入后超螺旋DNA的减少(SC%)与剂量呈良好线性关系,而γ射线和UV组的SC%随剂量升高呈指数下降;(3)干燥DNAγ辐照组的D37值为820Gy,SC完全消失的剂量LD为3814Gy,LD/D37=4.65;UV照射组的相应值分别为:1.65J/cm2,7.65J/cm2,4.64;N+离子组的相应值为:3.2×1015N+/cm2,5.0×1015N+/cm2和1.56。虽然上述三种辐射的剂量单位不同,不能直接比较其相对生物学效应,但从LD与D37的比值可反映DNASC破坏的程度和终点剂量(SC%=0)的大小。从而看出,N+离子(高LET辐射)比γ射线(低LET辐射)UV(非电离辐射)对DNA损伤作用更强;(4)乙醇、甘露醇等自由基清除剂对电离辐射损伤有很强的保护作用,但对UV损伤未见明显的保护效应  相似文献   

4.
观察血管紧张素Ⅱ(AngⅡ)对心肌肌浆网Ca2+,Mg2+-ATPase基因(SERCA2a)转录调节的影响,评价DMP811对此效应的干预作用.6周龄雄性SD大鼠随机分为3组,每组6只.组1:生理盐水输注;组2:AngⅡ输注+DMP811管饲(3mg·d-1·kg-1);组3:AngⅡ输注(200ng·min-1·kg-1.1周后称其体重,取心脏并称重,提取心脏总RNA后采用Northernblot的方法检测SER-CA2a的转录水平,采用RT-PCR检测AngⅡ1型受体(AT1)mRNA水平.实验后,组3心重(CW)、心重/体重(C/B)、AT1受体转录水平均高于组1(分别增加4.7±0.4%,4.9±0.9%和24.7±3.5%;P<0.01),而SERCA2a基因转录水平显著低于组1(降低20.1±3.0%,P<0.01),并且SERCA2amRNA水平与AT1受体mRNA水平呈负相关(r=-0.74,P<0.01).AngⅡ导致的上述改变能被DMP811完全阻断.AngⅡ通过其Ⅰ型受体的介导,诱导了SERCA2a的转录下调  相似文献   

5.
牡丹品种朱砂垒(Paeonia suffruticosa Andr.cv.Zhushalei)在冬季室内催花过程7种内源激素含量变化不同。玉米素核苷(Z+AR)、生长素(IAA)和赤霉素(GA3)的含量在花生长发 处于较高水平;而脱落酸(AB)、异戊烯基腺苷(IP+IPA)、二氢玉米素核苷(DHZ+DHZR)、赤霉素(GA4)的含量低于上述3种内源激素。激素平衡方面,GAs/ABA、CTKs/AB  相似文献   

6.
外源激素诱导风信子(Hyacinthus orientalisL.)同一发育时期花被外植体不同部位细胞再生花芽的实验表明∶1. 诱导花被外植体细胞再生花芽,外源激素是必需的;2. 仅有细胞分裂素就可以诱导花芽再生,生长素并不是必需的;3. 花被外植体上的不同部位的细胞再生花芽时,需要不同浓度的外源激素. 单独加6-BAP或玉米素2 m g/L可以诱导花被下部的细胞再生花芽;6-BAP或玉米素2 m g/L和2,4-D 0.1 m g/L的组合有利于花被中部的细胞再生花芽;6-BAP或玉米素2 m g/L和2,4-D 1.0 m g/L的组合能促进花被上部的细胞分化花芽  相似文献   

7.
用10μg/gLa ̄(3+)、1.0%Vc、10μg/gLa ̄(3+)+1·0%Vc、蒸馏水四种处理,喷施正处在衰老过程中的黄瓜叶片,一周后提取PSⅡ颗粒,通过SDS-PAGE、薄层扫描和Clark测氧等测试技术,发现处理与对照的PSⅡ颗粒在与放氧活性密切相关的33、23、17kD三肽的相对总含量上有变化,10μg/gLa ̄(3+)处理比对照减少28.9%,而1.0%Vc和10μg/gLa ̄(3+)+1.0%Vc处理的与对照接近。前者的放氧活性降低37.3%,后者的放氧活性增加62.4%,是对照的1.6倍,共同处理的结果与对照相近。可见,在光合放氧中,1.0%Vc与10μg/gLa ̄(3+)拮抗。而且,33、23、17kD三肽总量的相对减少与放氧活性的高低呈正相关。  相似文献   

8.
将含有鸡传染性支气管炎病毒 S1 基因c D N A 的重组转移质粒p S X I V V I+ X3 S1 . Holte 和p S X I V V I+ X3/4 S1 . Holte 分别与粉纹夜蛾核型多角体病毒 Tn N P V S V I- G D N A( O C C- ,gal+ ) 共转染草地夜蛾( Sf9) 细胞,经空斑纯化得到重组病毒 Tn N P V( X3) S1 . Holte O C C+ 和 Tn N P V( X3/4) S1 . Holte O C C+ 。将重组毒株分别感染 Tn5 B1 细胞,并进行 S D S P A G E 与 Westernblot 检测。结果表明, Tn N P V( X3/4) S1 . Holte O C C+ 在感染的细胞中高效表达了 S1 蛋白, S D S P A G E 凝胶薄层色谱分析结果显示,感染病毒后72 h S1 蛋白的表达量占细胞内总蛋白量的35 .8 % ,而 Tn N P V( X3) S1 . Holte O C C+ 感染的细胞内检测不出 S1 蛋白。经分析认为这一差异主要来自 S1 基因翻译起始位点及其附近的周围环境。  相似文献   

9.
目的和方法:本工作旨在研究免迷走复合区(DVC)内TRH对奥迪氏括约肌(SO)肌电的调节作用及外周途径。结果:(1)DVC内注射TRH(0.8nmol,1μl)后,慢波电位(SW)频率不变,但锋电位频率(FSPSO)及幅度(ASPSO)、锋电位发生率(ISP)明显增加.(2)DVC内分别注射不同剂量TRH(0.13,0.25,0.50,0.80,1.30nmol,1μl)后,各剂量TRH均能引起FSPSO增加。随注射剂量的增加,SO反应强度和持续时间均增加,呈现明显的剂量反应关系。(3)DVC内注射TRH兴奋FSPSO的效应可被静脉注射阿托品(0.2mg/kg)或迷走神经切断完全阻断,但不能被静脉注射酚妥拉明(1.5mg/kg)、心得安(1.5mg/kg)或脊髓切断术所阻断。结论:DVC内TRH可能对SO肌电有重要的调节作用,这种作用是通过迷走神经及外周M受体介导。  相似文献   

10.
目的 探讨将维生素A缺乏(VAD)胎鼠作为先天性心脏病动物模型的可行性。方法取11-19d不同胎龄正常及VAD胎鼠心脏经石蜡包埋、切片及 HE染色观察其发育情况。结果 1.实验组饲料含维生素A(VA)7μg/100g,经VAD饮食喂养后实验组大鼠血清VA水平明显低于对照组[(0.168±0.059)μmol/L Vs(2.18±0.23)μmol/L,t=32.88, P<0.001]。 2.大鼠死亡百分比:饲养于屏障系统的VAD大鼠死亡百分比较饲养于开放系统中的要低4.6倍(10% Vs 45.83%.x 2=16.64, P<0.001),对照组为0。 3.实验组大鼠受孕百分比及每只孕鼠产仔数均低于对照组[58.33% Vs 81.5%, x 2=4.37,P<0.05:(6.97±2.79) Vs(13 ±1.05),t=7.16, P<0.001]。 4.经切片观察11~15 d胎龄胎鼠实验组心脏出现明显发育延迟的占36.67%, 16~19 d胎龄胎鼠实验组心脏畸形占41.43%,血管异常占18.57%。结论VAD胎鼠可用来作为先天性心脏病动物模型,但需改进饲养环境以减少异常死亡。  相似文献   

11.
The major endogenous cytokinins, Z, ZR, DHZ, DHZR, iP and iPR in pistachio seedlings (Pistacia vera L. cv. Ohadi) were purified by HPLC and their identities confirmed using GC-MS. The aerial parts of two-year old pistachio seedlings including mature leaves, young leaves, lateral buds, debarked stems and bark were subjected to analysis. All of the above mentioned cytokinins were identified in the aerial parts except DHZ which was only present in mature leaves. Z-type cytokinins contributed almost 43% of the total cytokinins. ZR and DHZR were identified as the major ribosides and iP as the main base. The greatest concentration of ZR was detected in the bark, amounting to about 48%. DHZR and ZR constituted the major portion of the total cytokinins detected in both young and mature leaves while Z was detected as a minor cytokinin in leaves. The sharp increase of iP concentration during leaf maturation indicates that mature leaves are probably capable of de novo biosynthesis of cytokinins. The absence of DHZ (except in mature leaves) and the presence of considerable concentrations of DHZR in pistachio stems suggest that these tissues are able to metabolize DHZ to DHZR. The large amount of ZR in pistachio leaves suggests that root-derived ZR is transported into the leaves after loading into the xylem. The presence of high amounts of iP in pistachio lateral buds indicates that iP has been accumulated in these parts. The occurrence of a totally different cytokinin distribution pattern in buds, as compared with the other aerial parts, possibly results from their different metabolism.  相似文献   

12.
菊苣薄层培养花芽,营养芽分化中内源激素的动态变化   总被引:4,自引:0,他引:4  
菊苣(Cichorium intybusL.)花梗薄层细胞培养于MS附加NAA 和BA 或IAA 和BA 的MS培养基上有花芽或营养芽分化. 花芽分化中内源IAA、DHZ+ DHZR、iPA 含量明显增加,而Z+ ZR变化不明显.营养芽分化中内源细胞分裂素含量增加明显,而IAA 在培养前7 d 含量下降,随后有所增加,在原基形成时含量达原初水平的2/3. 可见,花芽分化比营养芽分化所需内源IAA/CTK 比值要高  相似文献   

13.
The sectioned thin cell layers (TCL) of flower stalk of Cichorium intybus L. were cultured in MS medium supplemented with NAA and BA or IAA and BA where floral and vegetative buds were developed from the explant. Endogenous IAA, DHZ+DHZR, iPA increased significantly during the floral bud formation, while Z+ZR remained changed. The levels of cytokinins, DHZ +DHZR, iPA, and Z-f-ZR all increased significantly during the vegetative bud formation, however IAA level was reduced during the first 7 days of culture and increased to two-thirds of initial values on the day when the bud primordia were formed. The results suggested that the initiation of floral buds was associated with a high IAA/CTK ratio, whereas the induction of vegetative bud differentiation was related to a low IAA/CTK ratio.  相似文献   

14.
High performance liquid chromatography analysis of immunoaffinity-purified extracts of mycelia of Amanita muscaria, and the Amaranthus bioassay of the eluted fractions, revealed the following seven cytokinins: zeatin, zeatin riboside, zeatin N-9-glucoside, dihydrozeatin, dihydrozeatin riboside, isopentenyl adenine, and isopentenyl adenosine. The decreased growth of aluminum-treated mycelia correlated with a 35% decrease in the total amount of the cytokinins. Among individual cytokinins, zeatin was the most affected, exhibiting a reduction of about 90%. The results are compared with previous investigations of aluminum effects on cytokinins in the mycelia of Lactarius piperatus, whose growth is stimulated by aluminum.Abbreviations ZR zeatin riboside - iPA isopentenyl adenosine - Z zeatin - DHZ dihydrozeatin - iP isopentenyl adenine - DHZR dihydrozeatin riboside - Z-9G zeatin N-9-glucoside - iP-9G isopentenyl N-9-glucoside - HPLC high performance liquid chromatography - DHZRMP dihydrozeatin riboside monophosphate - ZRMP zeatin riboside monophosphate  相似文献   

15.
Leaf senescence varies greatly among cotton cultivars, possiblydue to their root characteristics, particularly the root-sourcedcytokinins and abscisic acid (ABA). Early-senescence (K1) andlate-senescence (K2) lines, were reciprocally or self-graftedto examine the effects of rootstock on leaf senescence and endogenoushormones in both leaves and xylem sap. The results indicatethat the graft of K1 scion onto K2 rootstock (K1/K2) alleviatedleaf senescence with enhanced photosynthetic (Pn) rate, increasedlevels of chlorophyll (Chl) and total soluble protein (TSP),concurrently with reduced malondialdehyde (MDA) contents inthe fourth leaf on the main-stem. The graft of K2 scion ontoK1 rootstock enhanced leaf senescence with reduced Pn, Chl,and TSP, and increased MDA, compared with their respective self-graftedcontrol plants (K1/K1 and K2/K2). Reciprocally grafted plantsdiffered significantly from their self-grafted control plantsin levels of zeatin and its riboside (Z+ZR), isopentenyl andits adenine (iP+iPA), and ABA, but not in those of dihydrozeatinand its riboside (DHZ+DHZR) in leaves in late season, whichwas consistent with variations in leaf senescence between reciprocallyand self-grafted plants. The results suggest that leaf senescenceis closely associated with reduced accumulation of Z+ZR, andiP+iPA rather than DHZ+DHZR, or enhanced ABA in leaves of cotton.Genotypic variation in leaf senescence may result from the differencein root characteristics, particularly in Z+ZR, iP+iPA, and ABAwhich are regulated by the root system directly or indirectly. Key words: Abscisic acid, cotton, cytokinins, grafting, leaf senescence Received 23 October 2007; Revised 17 January 2008 Accepted 23 January 2008  相似文献   

16.
Ian M. Scott  Roger Horgan 《Planta》1984,161(4):345-354
the cytokinins of tobacco crown-gall tissue have been analysed by quantitative mass spectrometry using 2H2-labelled cytokinin riboside 5-monophosphates and 15N4-labelled cytokinin glycosides as internal standards. The principal endogenous cytokinin of this tissue is zeatin riboside 5-monophosphate. The biologically inactive 7-glucoside of zeatin is the most abundant basic cytokinin in the tissue. These findings expose the limitations of previously reported analyses of similar tissues, which were restricted to biologically active basic cytokinins. The present study demonstrates that the endogenous cytokinins of tobacco crowngall tissue show a clear correspondence to the range of metabolites formed when exogenous cytokinins are supplied to nontumorous tobacco cells.Abbreviations DHZ dihydrozeatin - DHZ7G dihydrozeatin 7-glucoside - DHZMP dihydrozeatin 9-riboside 5-monophosphate - DHZR dihydrozeatin 9-riboside - GC-MS coupled gas chromatography-mass spectrometry - HPLC high-performance liquid chromatography - Z7G zeatin 7-glucoside - Z9G zeatin 9-glucoside - ZOG zeatin O-glucoside - ZMP zeatin 9-riboside 5-monophosphate - ZR zeatin 9-riboside - ZROG zeatin 9-riboside O-glucoside  相似文献   

17.
T. L. Wang  R. Horgan 《Planta》1978,140(2):151-153
Dihydrozeatin riboside has been identified in the leaves of decapitated bean plants by Sephadex LH20 chromatography and combined gas chromatography-mass spectrometry. The relationship between the cytokinins isolated and identified from this system and those previously reported in Phaseolus is discussed.Abbreviations DHZ dihydrozeatin - DHZOG dihydrozeatin-O--D-glucoside - DHZR dihydrozeatin riboside - GCMS combined gas chromatography-mass spectrometry - GLC gas-liquid chromatography - TIC total ion current - TMS trimethylsilyl - Z zeatin - ZR zeatin riboside  相似文献   

18.
In this study, we report the role of exogenous and endogenous cytokinins on growth and sexual organ development in the fern Blechnum spicant L. Spore-derived gametophytes (SG) were cultured in full-strength Murashige and Skoog (1962) liquid medium supplemented with (a) 4.44 μMN6-benzyladenine (BAP), (b) a crude extract from mature female gametophytes, and (c) 4.44 μM BAP in combination with the crude extract from mature gametophytes, respectively. Both BAP and the crude extract delayed the gametophyte development, and this effect was increased when they were added together. With respect to sexual organ development, BAP inhibited the sexual organ formation, while the crude extract favored antheridia formation; however, when added together, the percentage of antheridia decreased. The endogenous level of the cytokinins cis-zeatin (cZ), cis-zeatin-riboside (cZR), dihydrozeatin (DHZ), dihydrozeatin riboside (DHZR), isopentenyl adenine (iP), isopentenyl adenosine (iPR), isopentenyl-9-glucoside (iP9G), trans-zeatin (tZ), and trans-zeatin riboside (tZR) were analyzed in female and male gametophytes of B. spicant L. The endogenous levels of cytokinins tZ, cZ, DHZ, cZR, iP, and iPR were higher in female gametophytes than in male gametophytes, with the endogenous iP and iPR content being increased more than 300 and 400 times, respectively.  相似文献   

19.
Flower stalk explants of tobacco cultured on a medium with an auxin and cytokinin regenerate flower buds within 14 days. The optimal medium concentrations of dihydrozeatin (DHZ) and benzyladenine (BA) were both 1 μM. The presence of DHZ in the culture medium was only essential during an initiation period of 7 days, whereas BA was needed only during the first 4 days. The difference in length of the initiation period is neither explained by the unequal uptake rates of the cytokinins nor by differences in their conjugation. At the medium concentration optimal for bud formation, the internal concentration of DHZ was two to three times the internal concentration of BA, which could be attributed to faster uptake of DHZ. It is concluded from the combined data that DHZ is less active in inducing flower bud formation than BA and that the exogenous cytokinins play only a role during the initiation phase of bud regeneration.  相似文献   

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